Abstract:
:Levels of acute phase cytokines secreted ex vivo by peripheral blood mononuclear cells (PBMCs) have been shown to be associated with clinical conditions or histologic lesions in renal transplant recipients. One of the limiting factors for the potential use of this assay as a diagnostic tool is the incubation time needed to measure adequate cytokine levels. Here, we validated that shorter time periods than the usual 48 h are sufficient for the production of acute phase cytokines. Cytokine levels were measured with the Luminex platform. We observed that, in contrast to cytokines associated with adaptive immunity, cytokines such as IL-1β, IL-6 and TNF-α are measurable as early as 2 h following incubation at a concentration of 1.5 million PBMC/150 μL. Levels obtained in the 2 h cultures have good correlations with the levels obtained after 48 h of culture for IL-1β and TNF-α (R=0.79, P=0.004 and R=0.92, P<0.001 respectively). We conclude that same-day incubation of PBMCs and measurement of these cytokines following blood collection in transplant recipients is feasible. It provides a rationale for further studies using shorter incubation times for ex vivo cellular assay measuring acute phase cytokine levels.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Mfarrej BG,Azzi J,De Serres SAdoi
10.1016/j.jim.2011.01.003subject
Has Abstractpub_date
2011-03-07 00:00:00pages
119-22issue
1-2eissn
0022-1759issn
1872-7905pii
S0022-1759(11)00006-8journal_volume
366pub_type
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